Usage of salmon sperm DNA in yeast transformation? - (Jun/26/2006 )

Hi all,

I am going to do transformation of a vector into pichia pasoris, but the protocol says must use salmon sperm DNA, so far I only know that it is used as blocking agent, but is it also used as blocking agent in the case of transformation? anyone knows what is it for? and how it works??

Thanks!

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ssDNA is used for carrier properties too when precipitating your ligation mix.

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i found a journal n a review about the usage of buffer n carrier dna (sperm dna) in yeast transformation. from thow references, i conclude the function of sperm dna in yeast transformation is to reduce membran binding to our plasmid. as we know, yeast cell is different with bacteria cell. in yeast, its membran can bind plasmid, so if we insert plasmid to yeast without addition of carrier dna, so we'll lost our plasmid in large number...

the review writer is Gietz.. n the journal by hinen...

i couldnt find gietz journal :-(

if you wanna the references you can contact me to w_ayu_hidayati@yahoo.com

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hi there,
You can read these articles. they will help you about purposes of usage of all chemical and biological components.
we performed yeast transformation in order to knock-out a gene, and we applied protocols in these articles. So, it works.

1-) High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method

R Daniel Gietz1 & Robert H Schiestl2

2-) Schiestl, R.H. & Gietz, R.D. High efficiency transformation of intact yeast cells using single-stranded nucleic acids as carrier. Curr. Genet. 16, 339–346 (1989).

3-)Gietz, R.D. & Woods, R.A. Genetic transformation of yeast. BioTechniques 30, 816–831 (2001).

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